Learning About Flow Cytometry and Cell Manipulation
Studying and manipulating cells in the laboratory is no easy task. First, scientists have to know what cells they will be manipulating, so that they know what tools they should use. Second, scientists should carefully establish their protocols, so that if anything goes wrong in the end processes following cell manipulation, they will know exactly what troubleshooting they need to do. Cell work begins with picking the right cell to work on, and this is why flow cytometry and cell manipulation are tied closely together.
The method of flow cytometry is used by laboratory and research scientists to count particles and sort them into different classes, as well as to examine them. During flow cytometry, cells are suspended in a fluid or liquid, and then passed through a detector that will sort and examine them at the same time. This detector usually employs a strong beam of light of a concentrated wavelength, and directs this onto the stream of liquid, where the cells are.
How Does Flow Cytometry Work?
All living cells and all particles will reflect, excite, absorb, or scatter light rays in a specific way, depending on the nature of their surfaces, their weight, and even their shape. Flow cytometers exploit this special property of solids by subjecting particles to several detectors in different positions with respect to the beam of light: one in line with the beam, and several other detectors perpendicular to it.
Some particles will scatter light in one manner, while others will scatter light in another manner; this is most obvious where fluorescent dies are concerned: some fluorescent chemicals can absorb light of one wavelength and emit or radiate it at another. Such properties are picked up by the flow cytometer’s detectors: through analysis of the detected brightness, it can be possible to come up with data on the various structures of the particles passing through the detectors, as well as important physical properties of the particles themselves.
In the case of flow cytometry in the life sciences, scientists can find out about the volume of a cell, the shape of its nucleus, the number of organelles, the nature of the cell membrane, and the presence of certain chemical compounds in the cytoplasm, all because of flow cytometry data. The most modern flow cytometers can analyze a few thousand particles each second, separate these particles or isolate them according to specific properties, and do all these tasks in real time. Flow cytometers are attached to computers, which scientists can use to analyze signals and look at data on the cells that they are studying.
The concepts of flow cytometry can be applied to other laboratory techniques, such as FACS, or fluorescence activated cell sorting. In this method, fluorescent dies can bind to certain cells, and these cells can be sorted out of the solution with the help of the flow cytometer. Flow cytometry can look at the pigments produced by cells, active protein or gene expression in certain cells, the expression of transgenes introduced to cells, enzyme activities, and the degree of cell death or apoptosis occurring in a cell population.
How Can Flow Cytometry be Used in Cell Manipulation?
There are thousands of applications of flow cytometry where the life sciences are concerned, and the applications mentioned above are the more important ones for cell manipulation. Flow cytometers can actually be used before and after cell manipulation. Scientists need viable and healthy cells to work with, and a flow cytometer can sort out healthy cells from a large population. Healthy cells may be those that have the correct morphology, or that are producing the right amount of enzymes that scientists need to study.
Cell manipulation can involve introduction of a transgene, or a gene that the cell does not normally have, but can express. For instance, E. coli cells are used to produce large amounts of insulin, and corn cells are induced to produce their own pesticides that will not harm humans. Not all cells in a manipulated population, however, can express a transgene correctly, and a flow cytometer can help sort the successfully manipulated cells out. Such a technique can also give scientists an idea of how efficient their manipulation methods are.
Cell manipulation might be clinical in nature: it can allow scientists to add compounds that can kill certain cells, such as cancer cells. Discovering such compounds can be valuable to the medical community; the more prospective cures there are for cancer, the better it will be for all of us. To look at the efficiency of such cures, scientists can add these compounds to cancer cells in vitro, and then use a flow cytometer to measure the degree of apoptosis, or cell death.
These are only a few applications of flow cytometry in cell manipulation. As research progresses in both these fields, we can be sure to see faster experimental methods, more efficient techniques in manipulating cells, and even faster flow cytometers that can carry out experiments in mere minutes. As technology advances, we can be sure to see more of flow cytometry and cell manipulation in the future.
